Investigation of Dimerization of G-alpha Protein by Föster Resonance Energy Transfer Method

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2018-09-05

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Son, Çağdaş Devrim

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The classical GPCR signaling pathway where, a heterotrimeric G proteinGPCR interaction is sufficient to transmit the signal to effector proteins, has been replaced by a heteromeric G proteinGPCR homo or heterodimer interaction model over the past two decades. These studies demonstrate that GPCRs that interact with each other or other GPCRs interact with a heteromeric G protein. In recent years, evidence suggest that dimer of GPCR dimers is required for some complex signal transductions.In these studies, it was proposed that this heteromeric tetramer formed by the dimerization of the dimers brought two G proteins close enough for proteinprotein interaction. It is not clear if GPCR tetramerization is required for Gprotein dimerization or dimerization can occure indipendent of the GPCRs. On the other hand, studies on small Gproteins (Ras family), which are structural homologs of G alpha subunits of heteromeric Gproteins, shown that dimerization can be independent of the receptors and necessary for various signaling pathways. Within the scope of this study, interactions of Gα proteins with each other were qualitatively and quantitatively investigated in live cells using bimolecular fluorescent complementation assay (BiFC) and Förster resonance energy transfer (FRET) methods. Results strongly suggest that GαiGαi interaction occures in N2a cell cultures. Findings of this study will help us understand the molecular mechanisms required for Gα dimerization and the dynamics of these proteins. Also, GPCR interactions with various effectors during complex signal transductions and the requirement of these receptors during Gα dimerizations can be studied with the techniques optimized in this study.

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https://hdl.handle.net/11511/81160

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Investigation of Gai1 protein homodimerization in live cells using förster resonance energy transfer (FRET) and biomolecular fluorescence complementation assay (BIFC) Atay, Özge; Son, Çağdaş Devrim; Department of Biochemistry (2019) The classical GPCR signaling pathway, where a heterotrimeric G protein-GPCR interaction is sufficient to transmit the signal to effector proteins has been replaced by a heteromeric G protein-GPCR homo- or hetero-dimer interaction model over the past two decades. These studies demonstrate that GPCRs that interact with each other couple with a heteromeric G protein. In recent years, evidence suggests that dimer of GPCR dimers is required for some complex signal transductions. In these studies, it was proposed...
INVESTIGATION OF PHYSICAL INTERACTION BETWEEN Gαi AND Gαs PROTEINS VIA FRET IN LIVE CELLS Balkan, Seyda Tuğçe; Son, Çağdaş Devrim; Küçük Baloğlu, Fatma; Department of Biochemistry (2021-8-11) GPCR’s are seven-transmembrane receptors that transmit external signals to the intracellular environment via secondary messenger systems through heterotrimeric G proteins. Heterotrimeric G proteins consist of α and β-γ subunits. Until recent years, scientists thought that GPCR signal transduction occurs between one GPCR and one heterotrimeric G protein; however, recently, it has been shown that GPCR’s can make oligomers. Oligomerization of GPCR allows cells to tune the intensity of the signal and respond ap...
Protein-Protein Interactions in Live Cells: Reinventing the Wheel Son, Çağdaş Devrim (2018-12-13) G protein-coupled receptors (GPCRs) are membrane proteins that mediate physiologicalresponse to a diverse array of stimuli. In humans, they mediate the action of hundreds ofpeptide hormones, sensory stimuli, odorants, neurotransmitters, and chemokines. GPCRs alsoare targets for ~40% of all currently marketed pharmaceuticals. These receptors traditionallybeen thought to act as monomeric units. However, recent evidence suggests that GPCRs mayform dimers as part of their normal trafficking and function. While ...
Visualization of interactions between fluorescently tagged g protein α11, α12/13 subtypes and adenosine 2a, dopamine 2 or homodimer adenosine 2a/2a receptors Kostromin, İrmak Begüm; Son, Çağdaş Devrim; Department of Biology (2018) G-Protein-coupled receptors (GPCRs) belong to one of the largest family of cell surface receptors, which transmit extracellular signals to intracellular responses by interacting with G- proteins. The G proteins are known as molecular switches that regulates different pathways via control of secondary messengers and signaling proteins. Adenosine 2A (A2A) and Dopamine 2 (D2) receptors belong to G-Protein-coupled receptors (GPCRs) family and are located mostly in striatopallidal γ-aminobutyric acid (GABA) cont...
Sequences in the intracellular loops of the yeast pheromone receptor Ste2p required for G protein activation. Celić, A; Martin, NP; Son, Çağdaş Devrim; Becker, JM; Naider, F; Dumont, ME (American Chemical Society (ACS), 2003-03-18) The α-factor receptor of the yeast Saccharomyces cerevisiae encoded by the STE2 gene is a member of the large family of G protein-coupled receptors (GPCRs) that mediate multiple signal transduction pathways. The third intracellular loop of GPCRs has been identified as a likely site of interaction with G proteins. To determine the extent of allowed substitutions within this loop, we subjected a stretch of 21 amino acids (Leu228−Leu248) to intensive random mutagenesis and screened multiply substituted alleles...

Citation Formats

Ç. D. Son, “Investigation of Dimerization of G-alpha Protein by Föster Resonance Energy Transfer Method,” 2018, Accessed: 00, 2021. [Online]. Available: https://hdl.handle.net/11511/81160.